Measuring the Infarct Zone and Brain Edema in a Rat Brain Following Middle Cerebral Artery Occlusion

Abstract

Source: Frank, D., et al. Measuring Post-Stroke Cerebral Edema, Infarct Zone and Blood-Brain Barrier Breakdown in a Single Set of Rodent Brain Samples. J. Vis. Exp. (2020).

This video demonstrates the assessment of the infarct zone and brain edema in brain slices from rats subjected to middle cerebral artery occlusion. Infarcted areas are identified using redox staining, while a blue filter and threshold function in image analysis software quantify the infarct zone and edema in the affected hemisphere.

Protocol

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.

1. Determination of infarct zone

1. Measure infarct zone (IZ) at 24 h after middle cerebral artery occlusion (MCAO).
   NOTE: Rats that lost more than 20% of their weight or developed seizures or hemiplegia are excluded from the experiment.
2. Euthanize the rat by replacing the inspired gas mixture with 20% oxygen and 80% carbon dioxide until the rat ceases to breathe spontaneously.
3. Open the chest with a 5-6 cm lateral incision through the abdominal wall ....

Results

Figure 1: Example scan of brain slices.

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
Brain & Tissue Matrices	Sigma-Aldrich	15013
Optical scanner	Canon	Cano Scan 4200F	Resolution 3200 x 6400 dpi
Petri dishes	Sigma-Aldrich	P5606
Scalpel blades #11	Sigma-Aldrich	S2771
Software
Adobe Photoshop CS2 for Windows	Adobe
ImageJ 1.37v	NIH		The source code is freely available. The author, Wayne Rasband (wayne@codon.nih.gov), is at the Research Services Branch, National Institute of Mental Health, Bethesda, Maryland, USA
Office 365 ProPlus	Microsoft	-	Microsoft Office Excel
Windows 10	Microsoft
Reagents
2,3,5-Triphenyltetrazolium chloride	Sigma-Aldrich	298-96-4
Evans blue 2%	Sigma-Aldrich	314-13-6